The results of our very successful phase I project showed that using projection lithography and off the shelf oligonucleotide synthesis reagents, we could in situ synthesize high density, high quality, very low cost, custom oligonucleotide microarrays on simple open substrates such as microscope slides. We also showed that using silver staining amplification it was possible to make microarrays which could be read by the eye and provide simple "yes" or "no" type of answers in resource poor environments. Phase II studies will focus on three specific aims: (i) Building a second generation custom microarray synthesizer which will incorporate all the knowledge we have gained in the past two years of operating our first generation synthesizer. The goal of this aim is to improve the quality of our custom microarrays even further while reducing cost, increasing density, flexibility and making them even easier to use by optimizing substrate surface chemistry. The second and third specific aims are for making new products for the next generation sequencers by taking advantage of the high quality and flexibility of our synthesis technology. In the second specific aim we will make custom arrays for capturing desired regions of genomic DNA by hybridization for targeted sequencing. The length of the oligonucleotides on the substrate surface and the capture and release protocols will be optimized to produce the highest selectivities possible. The third specific aim will develop a custom "RNA baits pool" kit for use in selecting desired regions of the genomic DNA in solution. We will take advantage of our high stepwise yields to produce long RNA baits to capture genomic DNA with efficiencies significantly higher than microarray hybridization based capture. Simultaneously with phase II research and development we will be commercializing the products of our technology which are ready for marketing (e.g. custom oligonucleotide microarrays) using our website based ordering system. While bringing in needed revenue with sales, we will look to increase our capital base by applying for a state of Michigan loan and get into discussions with venture capital investors to obtain the finances necessary for marketing on a larger scale. We already have several international distribution agreements and are in discussions with US companies with established marketing and distribution networks. By making custom high density custom microarrays, and selective custom capture kits for next generation sequencers much more affordable, our technology will have a significant impact on healthcare research and potentially in the diagnostic market. PUBLIC HEALTH RELEVANCE: This project aims to develop and commercialize high quality, low cost custom microarrays and genomic DNA capture kits. High density microarrays are used in a variety of research and diagnostic applications ranging from gene expression analysis, genotyping copy number variation to monitoring disease and treatment progression. Custom genomic DNA capture kits for next generation sequencers will enable researchers to study the regions of chromosomes they are most interested with much more detail and selectivity and speed up discoveries in the relating genetic information to disease.